Filtration-based systems and methods for isolation of clustered particles

Abstract

An embodiment of the disclosed technology provides an isolation device for isolating clustered particles. The isolation device can include an inlet configured to receive a fluid and an outlet configured to output the fluid. The fluid can include a plurality of non-clustered particles and a plurality of clustered particles. The isolation device can include a plurality of microwells. Each microwell can have a plurality of sidewalls and a bottom surfacing having a meshed trapping region. The meshed trapping region can capture the plurality of clustered particles while allowing the non-clustered particles to pass. The outputted fluid can include the plurality of non-clustered particle and be substantially free of the plurality of clustered particles.

1 . A system comprising:
 an isolation device formed by a fabricating process and comprising:
 an inlet; 
 microwells, each microwell including sidewalls and a bottom surface having a meshed trapping region; and 
 an outlet; 
 wherein the device is configured to:
 receive a fluid through the inlet comprising non-clustered particles and clustered particles; and 
 output an isolated fluid through the outlet comprising at least substantially all of the non-clustered particles and being at least substantially free of the clustered particles; 
 
 wherein the meshed trapping region is configured to capture at least substantially all clustered particles of the fluid and pass at least substantially all non-clustered particles of the fluid; and 
 wherein the meshed trapping region comprises apertures configured to divide a flow of the fluid into flow paths. 
   
     
     
         2 . The system of  claim 1  further comprising the fluid;
 wherein the fluid is blood, the non-clustered particles comprise non-clustered cells, and the clustered particles comprise cell-clusters. 
 
     
     
         3 . The system of  claim 1  further comprising the fluid;
 wherein the fluid is urine, the non-clustered particles comprise non-clustered cells, and the clustered particles comprise cell-clusters. 
 
     
     
         4 . The system of  claim 1 , wherein the device is further configured to provide a volumetric flow rate through the inlet and outlet of between approximately 20 mL/hour and approximately 100 mL/hour. 
     
     
         5 . The system of  claim 1 , wherein each microwell has a depth of between approximately 10 microns and approximately 500 microns. 
     
     
         6 . The system of  claim 1 , wherein at least a portion of each sidewall is slanted. 
     
     
         7 . The system of  claim 1 , wherein the device comprises between approximately 40 and approximately 280 microwells per millimeter squared. 
     
     
         8 .- 10 . (canceled) 
     
     
         11 . The system of  claim 1 , wherein the apertures are arranged in an array. 
     
     
         12 . The system of  claim 1 , wherein each aperture is sized such that the non-clustered particles pass through the apertures and the clustered particles do not pass through the apertures. 
     
     
         13 . The system of  claim 1 , wherein each aperture has a shape selected from the group consisting of a square, circle, ellipse, and polygon. 
     
     
         14 . The system of  claim 13 , wherein each aperture is square-shaped having a side length of between approximately 10 microns and approximately 17 microns. 
     
     
         15 .- 17 . (canceled) 
     
     
         18 . The system of  claim 1 , wherein each aperture has the same shape. 
     
     
         19 . The system of  claim 1  further comprising the fluid;
 wherein the clustered particles are label-free. 
 
     
     
         20 . The system of  claim 1  further comprising the fluid;
 wherein the clustered particles are labeled. 
 
     
     
         21 . The system of  claim 1 , wherein the device has a diameter of between approximately 5 millimeters and approximately 300 millimeters. 
     
     
         22 . The system of  claim 1 , wherein the device comprises a material selected from the group consisting of a fluorine-based polymer, a perfluoropolyether-based polymer, a heat-curable polymer, a UV-curable polymer, a metal, and a semiconductor. 
     
     
         23 .- 27 . (canceled) 
     
     
         28 . A fabricating process for the isolation device of  claim 1  comprising:
 fabricating a silicon mold on a silicon wafer; 
 fabricating a polymer mold; 
 fabricating the isolation device; and 
 releasing the isolation device. 
 
     
     
         29 . The fabricating process of  claim 28 , wherein fabricating the silicon mold on the silicon wafer comprises:
 depositing a first photoresist layer on the silicon wafer;   patterning the first photoresist layer;   etching the silicon wafer to form a plurality of pillars;   depositing a nitride layer on the silicon wafer;   depositing a second photoresist layer;   patterning the second photoresist layer and the nitride layer;   etching the silicon wafer to form slanted sidewalls extending to each pillar of the plurality of pillars;   depositing a third photoresist layer;   patterning the third photoresist layer; and   etching the silicon wafer to form the silicon mold.   
     
     
         30 . The fabricating process of  claim 28 , wherein fabricating the polymer mold comprises:
 coating the silicon wafer with silane;   depositing a first polymer layer on the silicon wafer;   curing the first polymer layer to form a first polymer mold;   removing the first polymer mold from the silicon wafer;   coating the first polymer mold with silane;   depositing a second polymer layer on the first polymer mold; and   curing the second polymer layer to form the second polymer mold.   
     
     
         31 . The fabricating process of  claim 30 , wherein the first polymer layer and the second polymer layer comprise polydimethylsiloxane (PDMS). 
     
     
         32 . The fabricating process of  claim 30  further comprising removing the second polymer mold from the first polymer mold. 
     
     
         33 . The fabricating process of  claim 30 , wherein fabricating the isolation device comprises:
 affixing the second polymer mold to a substrate;   filling the second polymer mold with a UV-curable polymer;   exposing the UV-curable polymer to UV light; and   curing the UV-curable polymer.   
     
     
         34 . The fabricating process of  claim 33 , wherein a vacuum pump is used to fill the second polymer mold with the UV-curable polymer. 
     
     
         35 . The fabricating process of  claim 33 , wherein the substrate is a vinyl dicing tape. 
     
     
         36 . The fabricating process of  claim 33 , wherein the substrate is an acetate sheet. 
     
     
         37 . The fabricating process of  claim 33 , wherein the substrate is a PET sheet. 
     
     
         38 . The fabricating process of  claim 33 , wherein filling the second polymer mold with the UV-curable polymer is performed on a thermoelectric cooler. 
     
     
         39 . (canceled) 
     
     
         40 . The fabricating process of  claim 33 , wherein releasing the isolation device comprises:
 removing the second polymer mold; and   removing the isolation device from the substrate.   
     
     
         41 . A method for isolating clustered particles using the system of  claim 1  comprising:
 passing a fluid through the isolation device, the fluid comprising clustered particles and non-clustered particles; 
 capturing the clustered particles within the meshed trapping region; and 
 outputting the isolated fluid comprising the non-clustered particles. 
 
     
     
         42 . The method of  claim 41 , wherein the fluid is blood, the non-clustered particles are cells, and the clustered particles are cell-clusters. 
     
     
         43 . The method of  claim 41 , wherein the fluid is urine, the non-clustered particles comprise non-clustered cells, and the clustered particles comprise cell-clusters. 
     
     
         44 . The method of  claim 41  further comprising positioning the isolation device within a filtration holder. 
     
     
         45 . The method of  claim 41 , wherein passing the fluid through the isolation device occurs at a flow rate of between approximately 20 mL/h and approximately 100 mL/h. 
     
     
         46 . (canceled) 
     
     
         47 . The method of  claim 41  further comprising retrieving at least a portion of the clustered particles from the meshed trapping region. 
     
     
         48 . The method of  claim 47 , wherein retrieving the clustered particles from the meshed trapping region comprises:
 washing the clustered particles with PBS; and   transferring the clustered particles to a holding container.   
     
     
         49 . The method of  claim 47 , wherein a micromanipulator retrieves the clustered particles directly from the meshed trapping region. 
     
     
         50 . The method of  claim 41  further comprising analyzing the clustered particles. 
     
     
         51 .- 52 . (canceled) 
     
     
         53 . The method of  claim 41  further comprising:
 coating the isolation device with a growth culture, wherein the captured clustered particles grow on the coated isolation device; and 
 analyzing the grown clustered particles directly on the coated isolation device. 
 
     
     
         54 .- 55 . (canceled) 
     
     
         56 . The method of  claim 41  further comprising coating the isolation device with a material selected from the group consisting of an inorganic material and an organic material. 
     
     
         57 .- 59 . (canceled) 
     
     
         60 . A method of detecting a clot using the system of  claim 1 . 
     
     
         61 . A method of dissociating a clustered particle using the system of  claim 1 .